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Multiple-stress is defined as the simultaneous or sequential exposure of plants to multiple abiotic constraints, which triggers regulatory programs that differ fundamentally from single-stress responses. In potato ( L.), drought, salinity, heat, and cold severely impair tuber development, yet the molecular architecture underlying resilience to combined stress remains unclear. We hypothesized that multi-stress conditions activate an integrated regulatory network linking tuber induction with stress-responsive metabolic and redox pathways. RNA-seq profiling of microtuberization under combined osmotic, salinity, heat, and cold stress was performed. Differential expression analysis identified shared differentially expressed genes (DEGs). A subset of upregulated genes was used for protein-protein interaction (PPI) network construction. Comparative regulatory analyses were performed, and selected genes were validated by qPCR. Statistical analyses were conducted to assess differential expression and network enrichment. A total of 2,046 shared DEGs were identified, including 1,212 upregulated and 834 downregulated genes. A PPI network constructed from 1,475 unique upregulated genes revealed 317 highly interconnected components. Network analysis identified the StSP6A-FD tuberigen complex as a central regulatory hub integrating developmental signaling with phenylpropanoid metabolism, oxylipin biosynthesis, and redox regulation. Multiple components were associated with hydrogen sulfide (H₂S) signaling, suggesting redox-gasotransmitter integration. Comparative regulatory analysis revealed conservation of the ERF-NAC-MYB-bZIP transcription factor framework, along with expansion of stress-responsive modules. Collectively, these findings establish a mechanistic framework linking tuber induction with adaptive metabolic remodeling under multi-stress conditions.

Wild Solanum species have contributed many introgressed genes during domestication into current cultivated potatoes, enhancing their biotic and abiotic stress resistance and facilitating global expansion. Abiotic stress negatively impacts potato physiology and productivity. Understanding the molecular mechanisms regulating tuber development may help solve this global problem. We made a transcriptomic analysis of potato microtuberization under darkness, cytokinins, and osmotic stress conditions. A protein–protein interaction (PPI) network analysis identified 404 genes with high confidence. These genes were involved in important processes like oxidative stress, carbon metabolism, sterol biosynthesis, starch and sucrose metabolism, fatty acid biosynthesis, and nucleosome assembly. From this network, we selected nine ancestral genes along with eight additional stress-related genes. We used qPCR to analyze the expression of the selected genes under osmotic, heat–osmotic, cold–osmotic, salt–osmotic, and combined-stress conditions. The principal component analysis (PCA) revealed that 60.61% of the genes analyzed were associated with osmotic, cold–osmotic, and heat–osmotic stress. Seven out of ten introgression/domestication genes showed the highest variance in the analysis. The genes H3.2 and GAPCP1 were involved in osmotic, cold–osmotic, and heat–osmotic stress. Under combined-all stress, TPI and RPL4 were significant, while in salt–osmotic stress conditions, ENO1, HSP70-8, and PER were significant. This indicates the importance of ancestral genes for potato survival during evolution. The targeted manipulation of these genes could improve combined-stress tolerance in potatoes, providing a genetic basis for enhancing crop resilience.

Purpose Bemisia tabaci ( Aleyrodidae family) is an insect vector of plant viruses that affects a wide variety of crops around the world. In the following study, we analyzed the variation in the bacterial communities associated with natural populations of B. tabaci (MEAM1) of four different crops from six regions from Mexico semi-arid zone (Baja California Sur). Methods PCR was used to amplify the mitochondrial cytochrome oxidase I gene (mtCOI), and then to carry out the phylogenetic analysis for genetic identification of the isolated B. tabaci. Next generation sequencing coupled with 16S metagenomic analysis was applied in order to characterize B. tabaci inner microbial community. Finally, bacterial obligate symbiont and facultative symbiont were confirmed by PCR amplification and by phylogenetic analysis. Results Ours results pointed toward that B. tabaci MEAM1 inner bacterial communities were predominantly structured by Proteobacteria phylum. Moreover, the most represented endosymbionts were the obligate endosymbiont from the genus “ Candidatus Portiera”, as well as two facultative symbionts belonging to genera Rickettsia and Hamiltonella ; both obligate and facultative endosymbionts were present for all samples, and their relative abundance varied was crop-independent. Conclusions Geographic localization and insect diet play a central role to maintain bacterial community structure of the B. tabaci MEAM1 whitefly at phylum taxonomic level. Agricultural practices were a factor that affected samples of bacterial community structure similarities, reflected in samples clustering. Host plants that are part of B. tabaci diet did not influence directly, in spite of sap nutrient differences, into obligate and facultative endosymbionts relative abundance.

Exposure of plants to stress conditions or to certain chemical elicitors can establish a primed state, whereby responses to future stress encounters are enhanced. Stress priming can be long-lasting and likely involves epigenetic regulation of stress-responsive gene expression. However, the molecular events underlying priming are not well understood. Here, we characterise epigenetic changes in tomato plants primed for pathogen resistance by treatment with β-aminobutyric acid (BABA). We used whole genome bisulphite sequencing to construct tomato methylomes from control plants and plants treated with BABA at the seedling stage, and a parallel transcriptome analysis to identify genes primed for the response to inoculation by the fungal pathogen, Botrytis cinerea . Genomes of plants treated with BABA showed a significant reduction in global cytosine methylation, especially in CHH sequence contexts. Analysis of differentially methylated regions (DMRs) revealed that CHH DMRs were almost exclusively hypomethylated and were enriched in gene promoters and in DNA transposons located in the chromosome arms. Genes overlapping CHH DMRs were enriched for a small number of stress response-related gene ontology terms. In addition, there was significant enrichment of DMRs in the promoters of genes that are differentially expressed in response to infection with B. cinerea . However, the majority of genes that demonstrated priming did not contain DMRs, and nor was the overall distribution of methylated cytosines in primed genes altered by BABA treatment. Hence, we conclude that whilst BABA treatment of tomato seedlings results in characteristic changes in genome-wide DNA methylation, CHH hypomethylation appears only to target a minority of genes showing primed responses to pathogen infection. Instead, methylation may confer priming via in-trans regulation, acting at a distance from defence genes, and/or by targeting a smaller group of regulatory genes controlling stress responses.

At present, the development of plants with improved traits like superior quality, high yield, or stress resistance, are highly desirable in agriculture. Accelerated crop improvement, however, must capitalize on revolutionary new plant breeding technologies, like genetically modified and gene-edited crops, to heighten food crop traits. Genome editing still faces ineffective methods for the transformation and regeneration of different plant species and must surpass the genotype dependency of the transformation process. Tomato is considered an alternative plant model system to rice and Arabidopsis, and a model organism for fleshy-fruited plants. Furthermore, tomato cultivars like Micro-Tom are excellent models for tomato research due to its short life cycle, small size, and capacity to grow at high density. Therefore, we developed an indirect somatic embryo protocol from cotyledonary tomato explants and used this to generate epigenetically edited tomato plants for the SlWRKY29 gene via CRISPR-activation (CRISPRa). We found that epigenetic reprogramming for SlWRKY29 establishes a transcriptionally permissive chromatin state, as determined by an enrichment of the H3K4me3 mark. A whole transcriptome analysis of CRISPRa-edited pro-embryogenic masses and mature somatic embryos allowed us to characterize the mechanism driving somatic embryo induction in the edited tomato cv. Micro-Tom. Furthermore, we show that enhanced embryo induction and maturation are influenced by the transcriptional effector employed during CRISPRa, as well as by the medium composition and in vitro environmental conditions such as osmotic components, plant growth regulators, and light intensity.

Coffee Coffea arabica L. depends on abundantly distributed rainfall, and drought negatively impacts plant development, fruit production, bean quality, and, ultimately, beverage quality. Plant biotechnology by means of genetic manipulation and plant regeneration by the somatic embryogenic process is an alternative technology to overcome these problems. In the present work, we used the molecular approach of the Trehalase gene silencing to allow trehalose accumulation favoring plants surviving in extreme drought/salt environments. We used a cassette containing the antisense C. arabica L. Trehalase gene under the RD29 promoter from A. thaliana and the NOS terminator to genetically modify an embryogenic coffee C. arabica L. cv Typica line under osmotic stress supplemented with mannitol (0.3 M) and sorbitol (0.3 M). Osmotic stress-tolerant somatic embryos lines were recovered and regenerated into plants. Tolerant somatic embryo lines showed a higher rate of competence to induce secondary SE capacity and plants robustness. These lines showed a down-regulation of the Trehalase; accumulation of trehalose, sucrose, starch, and proline; higher photosynthetic rate; improved water-use efficiency; and appropriated vapor deficit pressure under soil conditions. A transcriptome analysis was performed from highly competent somatic embryogenic lines to understand the molecular mechanisms underlying osmotic-stress tolerance. From the up-regulated genes, a PPI network made by STRING v12.0 with high confidence (0.700) revealed the presence of the 10 modules: the cell cycle, chromatin remodeling, somatic embryogenesis, oxidative stress, generic transcription pathway, carbon metabolism, phenylpropanoid biosynthesis, trehalose biosynthesis, proline biosynthesis, and glycerolipid metabolism.

Purpose The plant Solanaceae family is one of the most important for global agriculture and nutrition. Within this plant family, two plant species stand out for their economic importance and for human consumption, which are tomato ( Solanum lycopersicum ) and chili pepper ( Capsicum annuum ). Moreover, those plants support diverse and characteristic microbial communities that are uniquely suited to the plant habitat and intimately connected to plant health. The main objective of this work is the bacterial community characterization in the rhizobiome of tomato and chili pepper, cultivated in arid environments. Methods Five crop fields located in the south of the peninsula of Baja California, Mexico, were sampled. Total DNA was extracted from rhizosphere, rhizoplane, and endophytic root compartment and sequenced by Illumina MiniSeq platform technology applied to 16S rRNA gene V3 region. Results We were able to obtain 1,195,426 total reads and 1,725,258 total reads for tomato and chili pepper samples, respectively. The analysis of the bacterial community structures confirmed that the two plant species showed differences in their microbial community structures. Nonetheless, the microbial community structures were directly and equally influenced by the crop field localization of each plant species. Interestingly, we determined that in both plant species, the Proteobacteria was the main phylum. Conclusion In conclusion, we found that several bacterial families are part of the core rhizobiome (28 OTUs) for both tomato and chili pepper, but the most abundant were the Pseudomonadaceae family and the Pseudomonas genus, which most probably play a pivotal role in the microbial ecology to benefit both crop plants.

The common bean ( Phaseolus vulgaris L.) is the most important grain legume in the human diet with an essential role in sustainable agriculture mostly based on the symbiotic relationship established between this legume and rhizobia, a group of bacteria capable of fixing atmospheric nitrogen in the roots nodules. Moreover, root-associated bacteria play an important role in crop growth, yield, and quality of crop products. This is particularly true for legume crops forming symbiotic relationships with rhizobia, for fixation of atmospheric N 2 . The main objective of this work is to assess the substrate and genotype effect in the common bean ( Phaseolus vulgaris L.) root bacterial community structure. To achieve this goal, we applied next-generation sequencing coupled with bacterial diversity analysis. The analysis of the bacterial community structures between common bean roots showed marked differences between substrate types regardless of the genotype. Also, we were able to find several phyla conforming to the bacterial community structure of the common bean roots, mainly composed by Proteobacteria , Actinobacteria , Bacteroidetes , Acidobacteria , and Firmicutes . Therefore, we determined that the substrate type was the main factor that influenced the bacterial community structure of the common bean roots, regardless of the genotype, following a substrate-dependent pattern. These guide us to develop efficient and sustainable strategies for crop field management based on the soil characteristics and the bacterial community that it harbors.

Potato microtuber (MT) development through in vitro techniques are ideal propagules for producing high quality potato plants. MT formation is influenced by several factors, i.e., photoperiod, sucrose, hormones, and osmotic stress. We have previously developed a protocol of MT induction in medium with sucrose (8% w/v), gelrite (6g/L), and 2iP as cytokinin under darkness. To understand the molecular mechanisms involved, we performed a transcriptome-wide analysis. Here we show that 1715 up- and 1624 down-regulated genes were involved in this biological process. Through the protein–protein interaction (PPI) network analyses performed in the STRING database (v11.5), we found 299 genes tightly associated in 14 clusters. Two major clusters of up-regulated proteins fundamental for life growth and development were found: 29 ribosomal proteins (RPs) interacting with 6 PEBP family members and 117 cell cycle (CC) proteins. The PPI network of up-regulated transcription factors (TFs) revealed that at least six TFs–MYB43, TSF, bZIP27, bZIP43, HAT4 and WOX9–may be involved during MTs development. The PPI network of down-regulated genes revealed a cluster of 83 proteins involved in light and photosynthesis, 110 in response to hormone, 74 in hormone mediate signaling pathway and 22 related to aging.

The psyllid, , is an insect vector of ' Liberibacter' causing \"Zebra chip\" disease that affects potato and other crops worldwide. In the present study, we analyzed the bacterial communities associated with the insect vector central haplotype of tomato crop fields in four regions from Mexico. PCR was used to amplify the mitochondrial ( ) and then analyze the single nucleotide polymorphisms (SNP) and phylogenetic analysis for haplotype identification of the isolated . Moreover, we carried out the microbial diversity analysis of several collected from four regions of Mexico through the NGS sequencing of 16S rRNA V3 region. Finally, was detected by the gene PCR amplification, which is the facultative symbiont. Also we were able to confirm the relationship with several strains by phylogenetic analysis. Our results pointed that collected in the four locations from Mexico (Central Mexico: Queretaro, and Northern Mexico: Sinaloa, Coahuila, and Nuevo Leon) were identified, such as the central haplotype. Analyses of the parameters of the composition, relative abundance, and diversity (Shannon index: 1.328 ± 0.472; Simpson index 0.582 ± 0.167), showing a notably relatively few microbial species in . Analyses identified various facultative symbionts, particularly the ( : ) with a relative abundance higher. In contrast, the genera of and ' Carsonella' ( : : ) were identified with a relatively low abundance. On the other hand, the relative abundance for the genus ' Liberibacter' was higher only for some of the locations analyzed. PCR amplification of a fragment of the gene encoding a surface protein ( ) of and phylogenetic analysis corroborated the presence of this bacterium in the central haplotype. Beta-diversity analysis revealed that the presence of the genus ' Liberibacter' influences the microbiota structure of this psyllid species. Our data support that the members with the highest representation in microbial community of central haplotype, comprise their obligate symbiont, , and facultative symbionts. We also found evidence that among the factors analyzed, the presence of the plant pathogen affects the structure and composition of the bacterial community associated with .